Application of clusterization algorithms for analysis of semivolatile pollutants in Arkhangelsk snow.

The best way to understand the environmental status of a certain region involves thorough non-target analysis, which will result in a list of pollutants under concern. Arkhangelsk (64° 32' N 40° 32' E, pop. ~ 344,000) is the largest city in the world to the north of the 60th parallel. Several industrial enterprises and the "cold finger" effect represent the major sources of air contamination in the city. Analysis of snow with comprehensive two-dimensional gas chromatography-high-resolution mass spectrometry allows detecting and quantifying the most hazardous volatile and semivolatile anthropogenic pollutants and estimating long-term air pollution. Target analysis, suspect screening, and non-target analysis of snow samples collected from ten sites within the city revealed the presence of several hundreds of organic compounds including 18 species from the US EPA list of priority pollutants. Fortunately, the levels of these compounds appeared to be much lower than the safe levels established in Russia. Phenol and dioctylphthalate could be considered as the pollutants of concern because their levels were about 20% of the safe thresholds. ChromaTOF® Tile, MetaboAnalyst software platform, and open-source software protocols were applied to process the obtained data. The obtained clusterization results of the samples were generally similar for various tools; however, each of them had certain peculiarities. Bis(2-ethylhexyl) hexanedioate, benzyl alcohol, phthalates, aniline, dinitrotoluenes, and fluoranthene showed the strongest influence on the clusterization of the studied samples. Possible sources of the major pollutants were proposed: car traffic and pulp and paper mills.

Longitudinal association of prepubertal urinary phthalate metabolite concentrations with pubertal progression among a cohort of boys.

BACKGROUND: Epidemiological studies have reported associations of anti-androgenic phthalate metabolite concentrations with later onset of male puberty, but few have assessed associations with progression. OBJECTIVES: We examined the association of prepubertal urinary phthalate metabolite concentrations with trajectories of pubertal progression among Russian boys. METHODS: At enrollment (ages 8-9 years), medical history, dietary, and demographic information were collected. At entry and annually to age 19 years, physical examinations including testicular volume (TV) were performed and spot urines collected. Each boy's prepubertal urine samples were pooled, and 15 phthalate metabolites were quantified by isotope dilution LC-MS/MS at Moscow State University. Metabolites of anti-androgenic parent phthalates were included: butylbenzyl (BBzP), di-n-butyl (DnBP), diisobutyl (DiBP), di(2-ethylhexyl) (DEHP) and diisononyl (DiNP) phthalates. We calculated the molar sums of DEHP, DiNP, and all AAP metabolites. We used group-based trajectory models (GBTMs) to identify subgroups of boys who followed similar pubertal trajectories from ages 8-19 years based on annual TV. We used multinomial and ordinal regression models to evaluate whether prepubertal log-transformed phthalate metabolite concentrations were associated with slower or faster pubertal progression trajectories, adjusting for covariates. RESULTS: 304 boys contributed a total of 752 prepubertal urine samples (median 2, range: 1-6) for creation of individual pools. The median length of follow-up was 10.0 years; 79% of boys were followed beyond age 15. We identified three pubertal progression groups: slower (34%), moderate (43%), and faster (23%) progression. A standard deviation increase in urinary log-monobenzyl phthalate (MBzP) concentrations was associated with higher adjusted odds of being in the slow versus faster pubertal progression trajectory (aOR 1.47, 95% CI 1.06-2.04). None of the other phthalate metabolites were associated with pubertal progression. CONCLUSIONS: On average, boys with higher concentrations of prepubertal urinary MBzP had a slower tempo of pubertal progression, perhaps attributable to the disruption of androgen-dependent biological pathways.

Differentiation of Central Slovenian and Moscow populations of Rana temporaria frogs using peptide biomarkers of temporins family.

Skin secretion represents the only means of defense for the majority of frog species. That phenomenon is based on the fact that the main components of the secretion are peptides demonstrating greatly varying types of bioactivity. They fulfill regulatory functions, fight microorganisms and may be even helpful against predators. These peptides are considered to be rather promising pharmaceuticals of future generation as according to the present knowledge microorganisms are unlikely to develop resistance to them. Mass spectrometry sequencing of these peptides is the most efficient first step of their study providing reliably their primary structures, i.e., amino acids sequence and S-S bond motif. Besides discovering new bioactive peptides, mass spectrometry appears to be an efficient tool of taxonomy studies, allowing for distinguishing not only between closely related species, but also between populations of the same species. Application of several tandem mass spectrometry tools (CID, HCD, ETD, EThcD) available with Orbitrap mass analyzer allowed us to obtain full sequence of about 60 peptides in the secretion of Slovenian population of brown ranid frog Rana temporaria. The problem of sequence inside C-terminal cycle formed by two Cys and differentiation of isomeric Leu and Ile residues was done in top-down mode without any derivatization steps. Besides general biomarkers of Rana temporaria species, Central Slovenian population of Rana temporaria demonstrates six novel temporins and one brevinin 1, which may be treated as biomarkers of that population.

Changes in the Metabolism of Sphingoid Bases in the Brain and Spinal Cord of Transgenic FUS(1-359) Mice, a Model of Amyotrophic Lateral Sclerosis.

The aim of this study was to evaluate changes in the content of sphingoid bases - sphingosine (SPH), sphinganine, and sphingosine-1-phosphate (SPH-1-P) - and in expression of genes encoding enzymes involved in their metabolism in the brain structures (hippocampus, cortex, and cerebellum) and spinal cord of transgenic FUS(1-359) mice. FUS(1-359) mice are characterized by motor impairments and can be used as a model of amyotrophic lateral sclerosis (ALS). Lipids from the mouse brain structures and spinal cord after 2, 3, and 4 months of disease development were analyzed by chromatography/mass spectrometry, while changes in the expression of the SPHK1, SPHK2, SGPP2, SGPL1, ASAH1, and ASAH2 genes were assayed using RNA sequencing. The levels of SPH and sphinganine (i.e., sphingoid bases with pronounced pro-apoptotic properties) were dramatically increased in the spinal cord at the terminal stage of the disease. The ratio of the anti-apoptotic SPH-1-P to SPH and sphinganine sharply reduced, indicating massive apoptosis of spinal cord cells. Significant changes in the content of SPH and SPH-1-P and in the expression of genes related to their metabolism were found at the terminal ALS stage in the spinal cord. Expression of the SGPL gene (SPH-1-P lyase) was strongly activated, while expression of the SGPP2 (SPH-1-P phosphatase) gene was reduced. Elucidation of mechanisms for the regulation of sphingolipid metabolism in ALS will help to identify molecular targets for the new-generation drugs.

Cyclization of N-arylcyclopropanecarboxamides into N-arylpyrrolidin-2-ones under electron ionization and in the condensed phase.

RATIONALE: Mass spectrometry is known as an excellent method to predict the behavior of organic compounds in solution. The behavior of organic compounds in the gas phase inside the ion source of a mass spectrometer allows their intrinsic properties to be defined, avoiding the influence of intermolecular interactions, counter ions and solvent effects. METHODS: Arylpyrrolidin-2-ones were obtained by condensed-phase synthesis from the corresponding N-arylcyclopropanecarboxamides. Electron ionization (EI) with accurate mass measurements by high-resolution time-of-flight mass-spectrometry and quantum chemical calculations were used to understand the behavior of the molecular radical cations of N-arylcyclopropanecarboxamides and N-arylpyrrolidin-2-ones in the ion source of a mass spectrometer. The geometries of the molecules, transition states, and intermediates were fully optimized using DFT-PBE calculations. RESULTS: Fragmentation schemes, ion structures, and possible mechanisms of primary isomerisation were proposed for isomeric N-arylcyclopropanecarboxamides and N-arylpyrrolidin-2-ones. Based on the fragmentation pattern of the N-arylcyclopropanecarboxamides, isomerisation of the original M+• ions into the M+• ions of the N-arylpyrrolidin-2-ones was shown to be only a minor process. In contrast, this cyclization proceeds easily in the condensed phase in the presence of Brønsted acids. CONCLUSIONS: Based on the experimental data and quantum chemical calculations the principal mechanism of decomposition of the molecular ions of N-arylcyclopropanecarboxamides involves their direct fragmentation without any rearrangements. An alternative mechanism is responsible for the isomerisation of a small portion of the higher energy molecular ions into the corresponding N-arylpyrrolidin-2-one ions. Copyright © 2016 John Wiley & Sons, Ltd.

N-dealkylation of amines during water disinfection - Revealing a new direction in the formation of disinfection by-products.

Among numerous disinfection by-products (DBP) forming during aqueous chlorination nitrogen containing species are of special concern due to their toxicological properties. Nevertheless, corresponding reaction products of these natural and anthropogenic compounds are not sufficiently studied so far. An interesting reaction involves dealkylation of the substituted amine moiety. Here we present the results of the comparative study of one-electron oxidation and aqueous chlorination of several aliphatic and aromatic amines. The reaction products were reliably identified with gas chromatography - high resolution mass spectrometry (GC-HRMS), high pressure liquid chromatography - electrospray ionization high resolution mass spectrometry HPLC-ESI/HRMS), and electrochemistry - electrospray ionization high resolution mass spectrometry (EC-ESI/HRMS). Certain similarities dealing with the formation of the corresponding aldehydes and substitution of alkyl groups at the nitrogen atom for hydrogen were shown for the studied processes. The mechanism of the substituted amines' aqueous chlorination involving one-electron oxidation is proposed and confirmed by the array of the observed reaction products. Alternative reactions taking place in conditions of aqueous chlorination, i.e. aromatic electrophilic substitution, may successfully compete with dealkylation and produce major products.

Composition and antimicrobial activity of the skin peptidome of Russian brown frog Rana temporaria.

A nano-HPLC-ESI-OrbiTrap study involving HCD and ETD spectra has been carried out to clarify the composition of the skin peptidome of brown Russian frogs Rana temporaria. This approach allowed determinantion of 76 individual peptides, increasing 3-fold the identified portion of the peptidome in comparison to that obtained earlier with FTICR MS. A search for the new bradykinin related peptides (BRPs) was carried out by reconstructing mass chromatograms based on the ion current of characteristic b- and y-ions. Several peptides were reported in the secretion of R. temporaria for the first time. The overall antibacterial activity of the skin secretion in general and of one individual peptide (Brevinin 1Tb) was determined using PMEU Spectrion (Portable Microbe Enrichment Unit) technology. The inhibitory effects of these peptides on Staphylococcus aureus and Salmonella enterica Serovar typhimutium were equal in scale to that reported for some antibiotics.

Halogen substitution reactions of halobenzenes during water disinfection.

Although being successfully applied all over the world for more than 100 years water disinfection by means of chlorination possesses certain drawbacks, first of all formation of hazardous disinfection by-products (DBP). Aromatic halogenated DBPs significantly contribute to the total organic halogen and developmental toxicity of chlorinated water. The present study deals with investigation of possible substitution of one halogen for another in aromatic substrates in conditions of aqueous chlorination/bromination. The reaction showed high yields especially in case of substrates with proper position of an activating group in the aromatic ring. Thus, ipso-substitution of iodine by chlorine is the main process of aqueous chlorination of para-iodoanisole. Oxidation of the eliminating I+ ions into non-reactive IO3- species facilitates the substitution. Oxidation of eliminating Br+ is not so easy while being highly reactive it attacks initial substrates forming polybrominated products. Substitution of iodine and bromine by chlorine may also involve migration of electrophilic species inside the aromatic ring resulting in larger number of isomeric DBPs. Substitution of chlorine by bromine in aromatic substrates during aqueous bromination is not so pronounced as substitution of bromine by chlorine in aqueous chlorination due to higher electronegativity of chlorine atom. However, formation of some chlorine-free polybrominated products proves possibility of that process.

Mass spectrometric study of bradykinin-related peptides (BRPs) from the skin secretion of Russian ranid frogs.

Amphibian skin secretion is known to contain biologically active peptides. Bradykinins and related peptides (BRPs) can be found in these animals, while frogs from the genus Rana are considered to be leaders in the levels and variety of these peptides. A reasonable rationalization of this fact is that bradykinins are efficient defense compounds against predators. Forty-four various BRPs have been identified in the skin secretions of five ranid frog species (R. ridibunda, R. lessonae, R. esculenta, R. temporaria, R. arvalis) from the Zvenigorod region (Moscow district, Russia). Some of these peptides are already known, but the novel ones constitute a significant portion. An interesting group of novel peptides was isolated from R. lessonae. These are bradykinin analogues bearing a tyrosine residue in the 5th or 8th position. [Arg(0), Trp(5), Leu(8)]bradykinin and [Thr(6), Leu(8)]bradykinin that had been isolated from fish and avian species, respectively, were also detected in the frog secretion, supporting the predator defense hypothesis. Furthermore, a novel group of BRPs named 'lessonakinins' was discovered in R. lessonae and R. esculenta. All of them include the [Arg(0), Trp(5), Leu(8)]bradykinin sequence and have some structural resemblance to the precursor of this peptide cloned by Chen and coworkers recently. However, the C-terminal part of the lessonakinins does not match the sequence predicted by Chen, demonstrating possible incompleteness of information obtained by cDNA cloning.

GC-HRMS with Complementary Ionization Techniques for Target and Non-target Screening for Chemical Exposure: Expanding the Insights of the Air Pollution Markers in Moscow Snow.

Environmental exposure assessment is an important step in establishing a list of local priority pollutants and finding the sources of the threats for proposing appropriate protection measures. Exposome targeted and non-targeted analysis as well as suspect screening may be applied to reveal these pollutants. The non-targeted screening is a challenging task and requires the application of the most powerful analytical tools available, assuring wide analytical coverage, sensitivity, identification reliability, and quantitation. Moscow, Russia, is the largest and most rapidly growing European city. That rapid growth is causing changes in the environment which require periodic clarification of the real environmental situation regarding the presence of the classic pollutants and possible new contaminants. Gas chromatography - high resolution time-of-flight mass spectrometry (GC-HR-TOFMS) with electron ionization (EI), positive chemical ionization (PCI), and electron capture negative ionization (ECNI) ion sources were used for the analysis of Moscow snow samples collected in the early spring of 2018 in nine different locations. Collection of snow samples represents an efficient approach for the estimation of long-term air pollution, due to accumulation and preservation of environmental contaminants by snow during winter period. The high separation power of GC, complementary ionization methods, high mass accuracy, and wide mass range of TOFMS allowed for the identification of several hundred organic compounds belonging to the various classes of pollutants, exposure to which could represent a danger to the health of the population. Although quantitative analysis was not a primary aim of the study, targeted analysis revealed that some priority pollutants exceeded the established safe levels. Thus, dibutylphthalate concentration was over 10-fold higher than its safe level (0.001 mg/L), while benz[a]pyrene concentration exceeded Russian maximal permissible concentration value of 5 ng/L in three samples. The large amount of information generated during the combination of targeted and non-targeted analysis and screening samples for suspects makes it feasible to apply the big data analysis to observe the trends and tendencies in the pollution exposome across the city.

Novel natural peptides from Hyla arborea schelkownikowi skin secretion.

Hyla arborea schelkownikowi is one of the leaf frog species inhabiting the southern territories of Russia and the former USSR. This frog species is a member of the Hylidae Rafinesque, 1815 batrachians family. The present study deals with the previously uninvestigated peptidome of the Hyla arborea schelkownikowi skin secretion. Nano-electrospray ionization Fourier transform mass spectrometry (nanoESI-FTMS) of the skin secretion, in the intact form and after acetylation, was selected as the general method of analysis. Electron-capture dissociation (ECD) and collision-induced dissociation (CID) fragmentation were both employed, while de novo sequencing was performed by manual interpretation of the MS data. The suppression of the cyclization of b-ions in the mass spectrometer by the acetylation reaction proved to be very efficient for the de novo sequencing of short peptides. Ten skin peptides were found and all of them, except for bradykinin, had not previously been reported. Six of the peptides belong to the tryptophyllins and related peptides, while three peptides are similar to the aureins.

Arctic snow pollution: A GC-HRMS case study of Franz Joseph Land archipelago.

Anthropogenic pollution of the Arctic atmosphere is of great interest due to the vulnerability of the Arctic ecosystems, as well as the processes of global transport and accumulation of atmospheric aerosols at high latitudes under conditions of cold climate. The present work throws light upon chemical composition of Arctic snow as a natural deposition matrix for atmospheric semi-volatile pollutants taken from the northernmost Arctic archipelago - Franz Josef Land, which is least affected by local sources of pollution and being a unique unstudied environmental object. The used methodology involved the liquid-liquid extraction of snow samples with dichloromethane and combination of targeted and non-targeted analyses of semi-volatile organic compounds with comprehensive two-dimensional gas chromatography - high-resolution mass spectrometry. While almost none of the known priority pollutants (except three dialkylphthalates) were identified in the studied samples, non-targeted screening revealed a specific class of biomass burning biomarkers - fatty amides with oleamide being the major component among them. Some peculiar organic pollutants (N,N-dimethylcyclohexylamine and N,N-dimethylbenzylamine) were identified in few samples. First results on the semi volatile pollutants in Franz Joseph Land snow were obtained using the most reliable GC × GC-HRMS non-target analysis.

[Experimental study of proflavine acetate phototransformation processes].

Changes in proflavine acetate phototransformation processes upon exposure to visible-range irradiation were studied by high performance liquid chromatography. Proflavine acetate was offered as a photosensitizer during photodynamic water disinfection. Dye transformation products upon time-varying exposure to irradiation were identified. By using structure-activity relationships and information from toxicity databases, the authors evaluated the hazard of the identified products and identified the most hazardous ones.

[A mass-spectrometric approach to primary screening of collagenolytic enzymes].

A comparative analysis of MALDI TOF mass spectra of low-molecular products resulting from the hydrolysis of native collagen I by collagenases of various classes (bacterial metallocollagenase from Clostridium histolyticum, serine collagenase from the Morikrasa commercial preparation, cysteine collagenase from Serratia proteomaculans, and cysteine collagenases from larvae of beetles Dermestesfrischi and D. maculates) was carried out. The spectra contain a number of ion peaks common for all collagenases; nevertheless, the mass spectra of each hydrolysate contains a unique set of peaks ("fingerprint") characteristic of each enzyme. This is especially true for the peaks of major products with relative intensities of more than 50%. At the same time, the enzymes of one class (cysteine collagenases) exhibit in their mass spectra peaks of identical major products. The results show a potential opportunity for MALDI TOF application in the primary screening of collagenases according to the fingerprints of collagen hydrolysis products.

Regression algorithm for calculating second-dimension retention indices in comprehensive two-dimensional gas chromatography.

Gas chromatography-mass spectrometry (GC-MS) is one of the most accurate, well developed, and reliable analytical tools for the analysis of volatile and semivolatile compounds. The GC-MS data have been extensively improved by enhancing the separation capacity via comprehensive two-dimensional gas chromatography (GC × GC). The reliability of the identification of the analytes in GC × GC-MS can be notably improved by applying the second-dimension retention index (2I) as additional analytical parameter along with the commonly used first dimension retention index (1I) and mass spectrum. A novel approach for calculating second-dimension retention indices (2I) for semivolatile organic compounds is proposed. It is noteworthy that the standards used in calculations are the same compounds recommended as internal standards by US EPA 8270 Method for analysis of semivolatile organic compounds. The new algorithm takes into account the analyte retention time and its retention temperature at the secondary column, (2tR) and (2TR), respectively. The experimental data collected with different primary oven temperature ramp rates and carrier gas flow rates have shown that the calculated by the proposed approach 2I values remain the same for each evaluated compound, drifting in a very narrow range. The proposed approach was tested using 100 organic compounds from various chemical classes including alkanes, phenols, nitrobenzenes, chlorinated hydrocarbons, anilines, polycyclic aromatic hydrocarbons (PAHs), phthalates, etc. The important advantage of the proposed 2I values for compounds of the same chemical origin (reference standards and analytes) involves applicability of well-known Lee's indices for non-polar phases. Therefore, the proposed approach can be used in targeted and non-targeted analysis of a wide range of organic compounds. The reduced version of the second dimension retention indices provides a valuable mapping of the homologues series of organic compounds, making their detection and identification easy and reliable.

Detection of semi-volatile compounds in cloud waters by GC×GC-TOF-MS. Evidence of phenols and phthalates as priority pollutants.

Although organic species are transported and efficiently transformed in clouds, more than 60% of this organic matter remains unspeciated. Using GCxGC-HRMS technique we were able to detect and identify over 100 semi-volatile compounds in 3 cloud samples collected at the PUY station (puy de Dôme mountain, France) while they were present at low concentrations in a very small sample volume (<25 mL of cloud water). The vast majority (∼90%) of the detected compounds was oxygenated, while the absence of halogenated organic compounds should be specially mentioned. This could reflect both the oxidation processes in the atmosphere (gas and water phase) but also the need of the compounds to be soluble enough to be transferred and dissolved in the cloud droplets. Furans, esters, ketones, amides and pyridines represent the major classes of compounds demonstrating a large variety of potential pollutants. Beside these compounds, priority pollutants from the US EPA list were identified and quantified. We found phenols (phenol, benzyl alcohol, p-cresole, 4-ethylphenol, 3,4-dimethylphenol, 4-nitrophenol) and dialkylphthalates (dimethylphthalate, diethylphthalate, di-n-butylphthalate, bis-(2-ethylhexyl)-phthalate, butylbenzylphthalate, di-n-octyl phthalate). In general, the concentrations of phthalates (from 0.09 to 52 µg L-1) were much higher than those of phenols (from 0.03 to 0.74 µg L-1). To our knowledge phthalates in clouds are described here for the first time. We investigated the variability of phenols and phthalates concentrations with cloud air mass origins (marine vs continental) and seasons (winter vs summer). Although both factors seem to have an influence, it is difficult to deduce general trends; further work should be conducted on large series of cloud samples collected in different geographic areas and at different seasons.

Semi volatile organic compounds in the snow of Russian Arctic islands: Archipelago Novaya Zemlya.

Environmental contamination of the Arctic has widely been used as a worldwide pollution marker. Various classes of organic pollutants such as pesticides, personal care products, PAHs, flame retardants, biomass burning markers, and many others emerging contaminants have been regularly detected in Arctic samples. Although numerous papers have been published reporting data from the Canadian, Danish, and Norwegian Arctic regions, the environmental situation in Russian Arctic remains mostly underreported. Snow analysis is known to be used for monitoring air pollution in the regions with cold climate in both short-term and long-term studies. This paper presents the results of a nontargeted study on the semivolatile organic compounds detected and identified in snow samples collected at the Russian Artic Archipelago Novaya Zemlya in June 2016. Gas chromatography coupled to a high-resolution time-of-flight mass spectrometer enabled the simultaneous detection and quantification of a variety of pollutants including those from the US Environmental Protection Agency (EPA) priority pollutants list, emerging contaminants (plasticizers, flame retardants-only detection), as well as the identification of novel Arctic organic pollutants, (e.g., fatty acid amides and polyoxyalkanes). The possible sources of these novel pollutants are also discussed. GC-HRMS enabled the detection and identification of emerging contaminants and novel organic pollutants in the Arctic, e.g., fatty amides and polyoxyalkanes.

Electrospray ionization tandem mass spectrometry sequencing of novel skin peptides from Ranid frogs containing disulfide bridges.

Tandem mass spectrometry sequencing, as well as Edman sequencing of peptides belonging to the Rana genus, represents a difficult task due to the presence of a disulfide bridge at the C-terminus and their rather high molecular masses (over 2000 Da). The present study throws light upon the sequence of three rather long peptides (more than 20 amino acid residues each) isolated from the skin secretion of Russian frogs, Rana ridibunda and Rana arvalis. This novel aspect involves the fact that the sequences (including two sequences established de novo) were determined exclusively by means of mass spectrometry. A combination of electron capture dissociation (ECD) and collision-induced dissociaiton (CID) data accompanied by exact mass measurements (LTQ Fourier transform ion cyclotron resonance mass spectrometer) facilitated reaching the goal. To overcome the difficulty dealing with disulphide bridges ("Rana box"), reduction of the S-S bond with dithiotreitol followed by derivatization of Cys residues with iodoacetamide was used. The sequence was determined using combined spectral data on y and b series of fragment ions. A multiple mass spectrometry (MS(3)) experiment was also used to elucidate the sequence inside the "Rana box" after cysteine derivatization. Exact mass measurements were used to differentiate between Lys and Gln residues, while characteristic losses of 29 and 43 Da (d and w fragment ions) in CID and ECD experiments allowed us to distinguish between Ile and Leu isomeric acids.

[Products of the photolysis of 3,6-dichloropicolinic acid (the herbicide lontrel) in aqueous solutions].

We studied the composition of products of the photochemical degradation of 3,6-dichloropicolinic acid (DCPA), the active principle of Lontrel, a herbicide broadly used in agriculture. Ultraviolet irradiation (mimicking the natural sunlight action) did not degrade DCPA completely to environmentally safe products. The rate of DCPA degradation was notably lower when distilled water was replaced by river water and even lower in sea water. Chromatomass spectrometry revealed 9 compounds among the photolysis products, in addition to undegraded DCPA.

Molecular recognition of pseudodistamine isomeric precursors trans-3(4)-aminopiperidin-4(3)-ols by EI mass spectrometry.

Synthesis of drugs, biologically active compounds or their derivatives always requires precise and reliable method of their identification, including differentiation of the possible isomers. Pseudodistamines and their precursors became a matter of elevated attention due to their different enzymatic inhibition. This paper deals with one of the groups of the pseudodistamine precursors - trans-3(4)-aminopiperidin-4(3)-ols. Their synthesis brings to a mixture of 2 regioisomers, resulting in the necessity of their reliable recognition. NMR spectroscopy commonly used by organic chemists requires advance knowledge and experience to analyse the spectra of these regioisomers. Therefore, we herein proposed a simpler way to recognize trans-3(4)-aminopiperidin-4(3)-ols using mass spectrometry with electron ionization. Fragmentation of 4 pairs of aminopiperidinol regioisomers with variation of amine moiety was studied. The obtained results allowed defining a group of 3 ions ([M-18]+., [M-19]+, [M-43]+) related only to the structure of trans-4-aminopiperidin-3-ols and 1 ion (m/z 100) related to the structure of trans-3-aminopiperidin-4-ols. Besides, interrogation of intensity of ions common for spectra of both regioisomers allows making differentiation as well.